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Molecular Cloning and Expression of 6HIS - GFP Reporter Gene for Fluorescent and Electron Spectroscopic Imaging.

Published online by Cambridge University Press:  02 July 2020

M. Malecki ,
S. Sanchez ,
P. Skowron  and
G. Case
M. Malecki
Affiliation:
Integrated Microscopy Resource, National Institutes of Health Biotechnology Resource, University of Wisconsin at Madison, Madison, WI, USA. Laboratory of Molecular Biology, University of Wisconsin at Madison, Madison, WI, USA.
S. Sanchez
Affiliation:
Laboratory of Molecular Biology, University of Wisconsin at Madison, Madison, WI, USA.
P. Skowron
Affiliation:
Laboratory of Molecular Biology, University of Wisconsin at Madison, Madison, WI, USA.
G. Case
Affiliation:
Genetics Biotechnology Center, University of Wisconsin at Madison, Madison, WI, USA.
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Extract

Molecular cloning offers an opportunity for designing constructs for expressing chimeric proteins containing incorporated reporter molecules. In this approach, molecules are identified based upon reporter molecules which are expressed within cells as a result of transfection, instead of marking them through immunolabeling, in situ hybridization, or derivative incorporation. This approach is particularly useful for studies involving integrated microscopy. Integrated microscopy allows us to assemble images of the same cell obtained with different microscopes into one comprehensive message concerning cellular functions. Integration of fluorescence and electron spectroscopic imaging is particularly promising. The main advantage of this approach relies in overcoming limitations of each type of microscopy alone i.e. in studies on living cells limitations in spatial resolution of light microscopy and in analysis of supramolecular organization, limitation of electron microscopy to frozen or fixed cells. For this purpose, special reporter molecules suitable for selected modes of microscopy have to be used.

Type
Recent Advances in Labeling Techniques
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 994 - 995
Copyright
Copyright © Microscopy Society of America

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References

References:

1. This project is supported by the BIR-9522771 and BIR-9420056 grants from the NSF and the DRR- 570 grant from the NIH.Google Scholar
2. At present, Dr.Skowron, P. is at the University of Gdansk, Gdansk, PL.Google Scholar